OBJECTIVE To study the gene chip joint pyrosequencing technology in the newborn genetic deafness gene mutation screening, and provide a theoretical basis for the early diagnosis and prevention of genetic deafness. METHODS 2000 Neonatal EDTA umbilical cord blood was collected and genomic DNA (gDNA) was extracted. Microarray chip was used to detect four deafness gene at 9 mutation sites. And the positive result of gene chip detection was verified by pyrosequencing.RESULTS Among the GJB2 mutations, there were 1 case of 35delG mutation type, 3 cases of 176 del16 mutation type, 57 cases of 235del C mutation type, 9 cases of 299 del AT mutation type, 6 cases of GJB3 gene 538C>T mutation type. There were 5 cases of 1555A>G mutations and 1 case of 1494C>T mutations in mitochondrial 12S rRNA. There were 6 cases of 2168A>G mutation type and 23 cases of IVS7-2A>G mutations in SLC26A4. 103 cases of newborns carry the mutated gene in 2,000, the gene mutation rate is 5.15%. CONCLUSION All the four genes mutation at nine mutation sites are found in newborns with family history of non-hereditary deafness, and GJB2 gene mutation is common. The screening of genetic deafness in newborns is very important for early diagnosis and prevention of hereditary hearing loss. In particular, the diagnosis of mitochondrial 12S rRNA gene mutation can prevent the occurrence of deafness caused by drug use, for the genetic mutation of these carriers' health is of great significance.

OBJECTIVETo observe the effect platelet antigen modification by mPEG-SPA with different molecular masses.

METHODSPlatelet CD42a was modified by 5 kD and 20 kD mPEG-SPA, respectively, and the fluorescence intensity of CD42a was detect by flow cytometry and the three-dimensional structure of CD42a simulated to analyze the distribution of lysine in CD42a molecule.

RESULTSAfter platelet CD42a modification by 5 kD and 20 kD mPEG-SPA, the fluorescence intensity of CD42a decreased sharply by 85.54% and 88.65%, respectively, and multiple lysine regions were identified on the surface of CD42a molecule.

CONCLUSIONBoth 5 kD and 20 kD mPEG-SPA allow useful modification of platelet CD42a, but 20 kD mPEG-SPA is more advantageous than 5 kD mPEG-SPA.

Blood Platelets ; chemistry ; Humans ; Molecular Weight ; Platelet Glycoprotein GPIb-IX Complex ; chemistry ; Polyethylene Glycols ; chemistry ; Succinimides ; chemistry

Twelve compounds were isolated from Psoralea corylifolia and their structures were identified as isopsoralen (1), psoralen (2), 8-methoxypsoralen (3), psoralidin (4), corylin (5), bavachin (6), daidzein (7), corylifolinin (8), bavachinin (9), neobavaisoflavone (10), daidzin (11) and astragalin (12). The results showed that psoralidin had the activity of scavenging DPPH free radicals activity (IC50 43.85 mg x L(-1)). Psoralidin (IC50 1.32 mg x L(-1))c, oryfolin (IC50 4.97 mg x L(-1)), daidzin (IC50 10.47 mg x S(-1)), daidzein (IC50 34.22 mg) x L(-1)) and astragalin (IC50 31.27 mg x L(-1)) had the activity of scavenging ABTS free radical. Psoralidin (IC50 40.74 mg x L(-1)), coryfolin (IC50 45.73 mg x L(-1)) and daidzein (IC50 49.44 mg x L(-1)) had alpha-glucosidase inhibitory activity. Corylifolinin and neobavaisoflavone had significantly effect of inhibiting SA, MRSA and ESBLs-SA (MIC 0. 781 3, 1.562, 5, 0.781 25 microg x disc(-1) and 6.25, 6.25, 6.25 microg x disc(-1).
Anti-Infective Agents ; chemistry ; pharmacology ; Antioxidants ; chemistry ; pharmacology ; Bacteria ; drug effects ; Enzyme Inhibitors ; chemistry ; pharmacology ; Free Radical Scavengers ; chemistry ; pharmacology ; Glycoside Hydrolase Inhibitors ; Inhibitory Concentration 50 ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Psoralea ; chemistry

OBJECTIVETo discuss the mechanism of traditional Chinese medicines reducing phlegm and resolving masses in treatment of iodine deficiency-induced goiter by observing the expression of growth factors and the balance-regulating mechanism of proliferation and apoptosis.

METHOD180 four-week-old Wistar rats were selected to establish the iodine deficiency model. After the modeling, the rats were randomly divided into six groups: the normal control group, the model control group, the iodine group, the phlegm compound group, the L-T4 group and the phlegm compound and L-T4 group. At the 21st day and 77th day after administration, 15 rats in each group were killed to collect specimens. Doses were calculated and adjusted according to body surface area and body weight. TT3, TT4 radioimmunoassay, TSH, immunoradiometric method were adopted. Fas, FasL and PCNA protein expressions are detected using immunohistochemical methods.

RESULTCompared with the normal group and the model group, the expressions of fas and FasL in the phlegm Group significantly increased, the expressions of fas and FasL in the phlegm and L-T4 group were also increased significantly. The expression of fas in the L-T4 Group was significantly lower than that of the L-T4 group and the phlegm compound and L-T4 group. Compared with the normal group, the expression of PCNA of the phlegm group and the phlegm and L-T4 group was significantly lower. Compared with the model group, the expression of PCNA of the iodine group, the phlegm groups and the phlegm and L-T4 group were significantly lower. Compared with the normal group, the expression of VEGF in the iodine group significantly decreased after treatment. Compared with the iodine group, the expression of VEGF in the phlegm group and the L-T4 group significantly reduced. Compared with the normal group, the expression of TGF-beta1 in the model group and the phlegm group significantly increased. Compared with model group, the expression of TGF-beta1 in the iodine group significantly reduced. Compared with the phlegm group, the expression of TGF-beta1 in the phlegm compound and L-T4 group was significantly reduced.

CONCLUSIONTraditional Chinese medicines reducing phlegm and resolving masses can completely recover goiter by promoting apoptosis of thyroid cells, inhibiting their proliferation and the expression of growth factors and enhancing the expression of TGF-beta, without causing injury on thyroid cells.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Goiter ; drug therapy ; genetics ; metabolism ; Humans ; Male ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Rats ; Rats, Wistar ; Thyroid Hormones ; secretion ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the effect of a microRNA-132 antagonist on lithium-pilocarpine-induced status epilepticus (SE) in young Sprague-Dawley (SD) rats.

METHODSForty-five 3-week-old SD rats were randomly and equally divided into epilepticus model group, microRNA-132 antagonist group, and microRNA-132 antagonist negative control group. The young SD rat model of SE was established using lithium-pilocarpine. For the microRNA-132 antagonist group and the negative control group, pretreatment was performed 24 hours before the model establishment. Behavioral observation was performed to assess the latency of SE and success rate of induction of SE. The scale of Lado was used to evaluate the seizure severity. Electroencephalography (EEG) was used to assess the frequency and amplitude of epileptiform discharges. The mortality rate was calculated in each group.

RESULTSThere was no significant difference in the success rate of induction of SE between the three groups (P>0.05). Compared with the microRNA-132 negative control group and the epilepticus model group, the microRNA-132 antagonist group had significantly prolonged SE latency after model establishment (P<0.05), a significantly lower Lado score of seizure (P<0.05), significantly lower frequency and amplitude of epileptiform discharges on EEG (P<0.05), and a slightly reduced mortality rate.

CONCLUSIONSThe treatment with the microRNA-132 antagonist shows an inhibitory effect on the development and progression of lithium-pilocarpine-induced SE in young SD rats. The inhibition of microRNA-132 is likely to be a potential target or direction for drug treatment of SE.

Animals ; Electroencephalography ; Male ; MicroRNAs ; antagonists & inhibitors ; Pilocarpine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; drug therapy

OBJECTIVETo study the changes of body composition in females patients with human immunodeficiency virus (HIV)-related lipodystrophy (LD) syndrome (HIV-LD).

METHODSTotally 25 female patients who were treated in our hospital from January 2002 to December 2009 were divided into LD group and non-LD group based on the existence of LD. All these patients were receiving highly active antiretroviral therapy (HAART). In addition, 12 healthy women were set as the controls. Total and regional body composition were measured by dual X-ray absorptiometry in all three groups.

RESULTSThe fat mass (FM) was correlated negatively with the duration of HAART (r=-0.431, P=0.029). Multiple linear regression analysis showed that FM had positive correlation with weight and negative correlation with lean mass (LM) (r = - 0. 973, P =0. 000). Total, trunk and leg FM were significantly lower in LD patients than that in controls (P <0.05).Meanwhile, total, trunk and leg bone mineral contents were statistically lower in LD patients than that in controls (P <0. 05). Lumbar bone mineral density of LD patients was lower than that of non-LD patients and controls, and there was significant difference between LD patients and controls (P = 0. 001). LM of LD patients was higher than that of non-LD patients but without statistical difference (P > 0. 05).

CONCLUSIONSThe peripheral and central FM and bone mineral contents remarkably decrease in female patients with HIV-LD. How-ever, HIV-LD patients tend to have higher LM than non-LD patients. .

Adipose Tissue ; metabolism ; Adult ; Body Composition ; physiology ; Bone Density ; physiology ; Female ; HIV-Associated Lipodystrophy Syndrome ; metabolism ; Humans ; Middle Aged ; Young Adult

OBJECTIVETo observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.

METHODSForty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.

RESULTS(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).

CONCLUSIONThe moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.

Acupuncture Points ; Animals ; Arthritis, Experimental ; metabolism ; therapy ; Disease Models, Animal ; Female ; Humans ; Moxibustion ; Potassium ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the additional efficacy of local anesthetic injection (LAI) as a part of multimodal anal?gesia in patients undergoing total knee arthroplasty (TKA) with respect to pain, narcotic use, knee function and complications. Methods A multicenter randomized, controlled, double blind study was performed. A total of 101 patients undergoing unilateral TKA in two centers were randomly divided into injection group and control group. Injection group (50 cases) received local anes?thetic injection of ropivacaine (200 mg), fentanyl (1μg) and epinephrine (1∶1 000, 0.25 mg) in operation and control group (51 cas?es) did not. All patients received standardized general anesthesia and postoperative intravenous patient controlled analgesia (PCA). Preoperative baseline data, surgery?related conditions, postoperative pain (on a 0 to 10 scale), knee function, time of open?ing PCA, narcotic dosage in PCA and complications were compared respectively. Results The time of opening PCA in injection group (4-10 h, M=8 h) was longer than that in control group (2-5 h, M=4 h) (P<0.05). The 12 h, 24 h and total narcotic use of PCA in injection group (8.62±3.601 ml, 21.22±9.220 ml, 38.52±7.764 ml) was less than that in control group (18.43±9.671 ml, 35.30± 11.414 ml, 55.52±12.405 ml) (P<0.05). At post anesthesia care unit the mean VAS in injection group (2.40±1.927) was lower than that in control group (3.06 ± 2.073) (P<0.05). There was no difference in mean VAS at other time points, knee function, length of stay between two groups (P>0.05). Conclusion LIA in TKA can relieve pain early after TKA, prolong the time of opening PCA and reduce narcotic use compared with patients without it. It is simple and safe to use.

OBJECTIVETo evaluate the influence of different pneumoperitoneal media on colon carcinoma LS-174T cell proliferation in vitro.

METHODSThe artificial pneumoperitoneum was established. The proliferation of LS-174T cells was detected by MTT assay and soft agar clone formation assay. Expression of HIF-1alpha and VEGF was examined by immunohistochemistry. Apoptosis of LS-174T cells was analyzed by AO/EB double fluorescein stain and flow cytometry.

RESULTSThe growth speed and proliferating capacity of LS-174T cells in CO(2) pneumoperitoneum group[A:0.37 +/- 0.02,formation (32.8 +/- 3.6)%] were significantly higher than those in control group [A:0.33 +/- 0.01,formation (28.4 +/- 2.3)%] and He group [A:0.30 +/- 0.01,formation (23.5 +/- 2.7)%], meanwhile the He group was the lowest (P<0.01). Positive expression of HIF-1alpha and VEGF in CO(2) and He artificial pneumoperitoneum up-regulated significantly as compared to control group(P<0.01), meanwhile the above expression was higher in CO(2) group (P<0.01). The G(0 )/G(1) ratio in CO(2) group was the lowest as compared to control group and He group (P<0.01), and G(0 )/G(1) ratio in He group was higher than that of control group(P<0.01). Aapoptosis rate in He group was the highest as compared with the other two groups(P<0.01).

CONCLUSIONCO(2) pneumoperitoneum has stronger effect on the proliferation of colon carcinoma cell LS-174T as compared to He pneumoperitoneum in vitro.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; pathology ; Flow Cytometry ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Pneumoperitoneum, Artificial ; methods

Using naproxen as model drug, the formulation of microemulsion vehicle for transdermal delivery was optimized by genetic algorithm. The ranges of microemulsion composed of Tween 80, IPM, alcohol and water, were defined through construction of the pseudo-ternary phase diagrams. Systematic model microemulsions containing naproxen 1.12% were prepared by a three-factor, three-level center design method, and their diffusion studies via excised rabbit skin were performed. Using the quadratic regression model of steady-state permeation rate (Js) of naproxen as objective function, the consequence of center design experiment was optimized by genetic algorithm, and the formulation of microemulsion with highest permeation rate for naproxen was screened. The optimum formulation was composed of 21.41% Tween 80, 15.17% alcohol, 4.14% IPM, and 59.28% water, and the anticipated Js was 183.57 microg x cm(-2) x h(-1). The results of back substitution test showed the steady-state permeation rate of naproxen microemulsion prepared according to optimum formulation was 189.43 microg x cm(-2) x h(-1), which was higher than anticipated value. This result demonstrated optimizing formulation of microemulsion for transdermal delivery by genetic algorithm is feasible, reliable and reasonable.
Administration, Cutaneous ; Algorithms ; Animals ; Models, Genetic ; Naproxen ; administration & dosage ; pharmacokinetics ; Rabbits ; Skin Absorption